Introduction: Fasting during Ramadan is a religious obligation for healthy adult Muslims. This study aimed to investigate the effects of Ramadan fasting and physical activity on blood hematological and biochemical parameters in male and female subjects. Methods: In this study, 50 healthy subjects were randomly assigned to four groups: male control fasting (MCF) group (n=13), male training fasting (MTF) group (n=13), female control fasting (FCF) group (n=12), and female training fasting (FTF) group (n=12), and were compared in two stages (before and after the intervention). For inter- and intra-group comparisons, a repeated measure ANOVA was applied. Results: The results showed a significant decline in body weight, body mass index, body fat percentage, and waist-to-hip ratio after Ramadan fasting, compared to the pre-Ramadan period. Also, significant changes were observed in red blood cell count and hematocrit level (P<0.05). Additionally, serum total cholesterol (TC), low-density lipoprotein (LDL), and TC/high-density lipoprotein (HDL) ratio significantly decreased after Ramadan, compared to the pre-Ramadan period. The two groups were significantly different in terms of weight, body mass index, body fat percentage, and waist-to-hip ratio after Ramadan (P<0.05). Moreover, the two groups were significantly different in terms of changes in mean triglyceride level, TC concentration, LDL-cholesterol, LDL/HDL ratio, TG/HDL ratio, and TC/HDL ratio. Conclusion: Ramadan fasting, accompanied by regular physical activity, can induce positive changes in hematological and biochemical parameters. These variations may be due to dietary changes, biological responses of the body to starvation, or physical activity during Ramadan.